80203 Monoclonal Anti cAMP EIA Kit

Monoclonal Anti-cAMP Antibody Based
Direct cAMP ELISA Kit
（New Non-acetylated Version）
Catalog No. 80203
96 Well Kit

Product Description
Adenosine 3', 5'-cyclic monophosphate （cyclic AMP; cAMP） modulates various physiological functions
such as cardiovascular biology, learning and memory, olfaction, immune response, asthma and kidney
function （1,2）. cAMP is produced from ATP by adenylyl cyclases and is degraded by
phosphodiesterases. Stimulation of adenylyl cyclases or inhibition of phosphodiesterases can increase
cellular cAMP concentrations. Blockers of adenylyl cyclase-activating receptors and inhibitors of the
cAMP-specific phosphodiesterases are used for treating human diseases. For example, blocking agents
for cAMP-increasing beta-adrenergic receptors （beta-blockers） are used for
treating abnormal heart rhythms, high blood pressure （hypertension）,
myocardial infarction and heart failure. Inhibitors of cAMP specific
phosphodiesterase types 2 and 4 are being tested for cognition
enhancement.
To screen for inhibitors or stimulators of cellular cAMP levels, it is
essential to have a sensitive, selective and reproducible method to measure
the cAMP concentrations. This is especially true for the initial screenings
given the possible weaker effects of larger pools of compounds.
Currently available other ELISA kits measuring cAMP levels are based on
the non-affinity-purified polyclonal anti-cAMP antibody. Despite the
claimed selectivity, these polyclonal anti-cAMP antibodies display certain
cross-reactivity with ATP. Given that ATP is the substrate for the cAMP production, it is very desirable
to have an antibody with high specificity towards cAMP over ATP.
NewEast Biosciences cAMP ELISA kit is based on the unique mouse monoclonal anti-cAMP
antibody. This monoclonal anti-cAMP antibody displays >108 fold of selectivity over ATP, cGMP, and
other nucleoside analogues. NewEast Biosciences cAMP ELISA kit provides significantly improved
sensitivity and selectivity over other kits based on polyclonal anti-cAMP antibodies. Our monoclonal
anti-cAMP antibody-based ELISA kit also avoids the batch-to-batch variations associated with
polyclonal antibody productions from animals, thus providing the reproducibility in the long run.
Furthermore, while polyclonal anti-cAMP antibodies used in other ELISA kits have higher affinity for
acetylated cAMP than non-acetylated cAMP, NewEast Biosciences monoclonal anti-cAMP antibody has
similar affinities to non-acetylated and acetylated cAMP molecules. Therefore, acetylation treatments of
samples and standards are not needed in NewEast Biosciences cAMP ELISA kit. This significantly
reduces the time for the assay. The avoidance of organic reagents used in the acetylation process
provides a safe and healthy work environment.
Principle Outline
NewEast Biosciences cAMP ELISA Kit is a competitive immunoassay to measure the cAMP levels,
either from cell extracts or from in vitro adenylyl cyclase assays. Briefly, multi-well plates are coated
with goat-anti-mouse serum. cAMP in cell extracts or in in vitro adenylyl cyclase assays will
competitively bind to the monoclonal anti-cAMP antibody in the presence of fixed amounts of
cAMP-conjugated horse-radish peroxidase or alkaline phosphatase. Known amounts of cAMP are used
as standards to generate the calculation curve. After a short incubation, the excess reagents are washed
away and substrate is added. The multiwell plates are then read on a microplate reader at 450 nm or 405nm. The intensity of the yellow color is inversely proportional to the concentration of cAMP in samples.
The measured optical density is used to calculate the concentration of cAMP in samples based on the curve
from the cAMP standards.